Cationic liposome enhancement of adenoviral vectors for gene delivery to OSCC

ORCiD

Nejat Düzgüneş: 0000-0001-6159-1391

Department

Biomedical Sciences

Document Type

Conference Presentation

Conference Title

39th Annual Meeting of the American Association for Dental Research (AADR)

Location

Washington, DC

Conference Dates

March 3-6, 2010

Date of Presentation

3-5-2010

Journal Title

Journal of Dental Research

Journal ISSN

0022-0345

Journal Volume Number

89 (Special issue A)

First Page

1134

Abstract

Objectives: To test the hypotheses that (i) adenoviral vectors will efficiently deliver the tumor-specific, survivin-driven luciferase gene to oral squamous cell carcinoma (OSCC) cells, and (ii) cationic liposomes will enhance adenoviral transduction. Methods: Two recombinant adenoviral vectors that encode luciferase were used: Ad-Sur-luc and Ad-CMV.luc with the human survivin and cytomegalovirus promoters, respectively. A recombinant adenovirus with polylysine-modified fiber knobs (Ad-pK7-CMV.luc) was also tested. The viruses were incubated with HSC-3, H413, and H357 human OSCC cells at different MOI, and luciferase expression was determined after 48 h. The viruses were also mixed with Metafectene or Metafectene-Pro (Biontex), before incubation with the cells to assess the effect of cationic liposomes on transduction. Results: Representative luciferase activities with Ad-CMV.luc (MOI 100) in HSC-3, H413 and H357 cells were 30460±1510, 10305±529 and 30907±1015 RLU/ml, respectively. Luciferase activities obtained with Ad-Sur.luc were 200±36, 97±12 and 100±8 RLU/ml, respectively. Metafectene Pro complexed with Ad-CMV.luc increased gene expression in all cell lines by 4 to 9-fold. Metafectene Pro complexed with Ad-Sur.luc enhanced luciferase expression by 13 to 17 fold. Ad-pK7-CMV.luc incubation with HSC-3, H413 and H357 cells resulted in luciferase activities of 15081±1694, 14427±3099 and 24187±272 RLU/ml, which decreased to 4037±446, 3359±206 and 8017±254 RLU/ml, respectively, when the virus was pre-incubated with Metafectene-Pro. Conclusion: Transduction of OSCC cells with adenoviral vectors alone did not result in high levels of reporter gene expression that were expected. However, complexing cationic liposomes with the viral vectors greatly improved gene expression. The use of viral vectors in conjunction with cationic liposomes can be a promising tool to help achieve the delivery of suicide genes in the therapy of OSCC. Supported by a Research Pilot Project Award (DRES03-Activity 062) from the University of the Pacific, Arthur A. Dugoni School of Dentistry (J.O.).

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